Note Antidepressant-Like Effects of an Aqueous Extract of Lavender

Food Sci. Technol. Res., 18 (3), 473–479, 2012 Note

Antidepressant-like Effects of an Aqueous Extract of Lavender

(Lavandula angustifolia Mill.) in Rats

1, 3 2 1 2 1 1, 4* Aya Kageyama , Toshio Ueno , Mutsumi Oshio , Hideki Masuda , Hiroko Horiuchi and Hidehiko Yokogoshi

1 Laboratory of Nutritional Biochemistry and Global COE Program, Graduate School of Nutritional and Environmental Sciences, Univer- sity of Shizuoka, Shizuoka 422-8526, Japan 2 Laboratory of Health Materials, Functional Ingredient Department, Ogawa & Co., Ltd., 15-7 Chidori Urayasushi, Chiba 279-0032, Japan 3 Present address: Division of Medical Nutrition, Tokyo Healthcare University, Tokyo 154-8568, Japan 4 Present address: Department of Food & Nutritional Sciences, Chubu University, Aichi 487-8501, Japan

Received June 30, 2011; Accepted January 5, 2012

The purpose of the present study was to examine the antidepressant-like effects of an aqueous extract of lavender (LAE) using the forced swimming test (FST), the most widely used animal model of depression. LAE was orally administered to rats three times, i.e., 24, 5, and 1 h prior to the FST. The administration of LAE (3428 mg/kg body weight) showed a significant reduction of the immobility time in the FST, the effect of which was comparable to that of the synthetic antidepressant, imipramine (30 mg/kg). In addition, the same dose of LAE did not change the locomotor activity in the open field test. These results suggest that LAE might have antidepressant-like effects that are independent of motor stimulation. Fur- thermore, the active ingredients of LAE were suggested to be non-volatile constituents, because linalool, the main aroma constituent of lavender, was completely removed during the preparation of LAE. Possible contribution of rosmarinic acid and that of apigenin glycosides to the antidepressant-like effects of LAE were discussed on the basis of their content in LAE.

Keywords: lavender, depression, antidepressant-like, forced swimming test

Introduction Lavender (Lavandula angustifolia Mill.) is a famous herb Depression has become a common psychological ill- that has a long history in folk medicine and is still therapeu- ness in recent years. According to an investigation by the tically used today. The essential oil obtained by steam distil- World Health Organization International Consortium of lation from the fresh flowering tops of this plant is often used Psychiatric Epidemiology (WHO-ICPE), 6.3 − 15.7% of in aromatherapy as a relaxant (Lis-Balchin, 1996). Inhalation the world’s population has been estimated to get depression of the vapor of the lavender essential oil and its main con- once in their life (Andrade et al., 2003). Exposure to chronic stituent, linalool, has shown sedative effects in both human stress is known to be associated with a higher incidence of and animal studies (Buchbauer et al., 1991; Buchbauer et depression, though its mechanism has not been elucidated. al., 1993; Diego et al., 1998; Kuroda et al., 2005). Other Although a wide variety of antidepressant drugs are avail- pharmacological effects of this oil, including anticonvulsive able to treat depression, most of the synthetic drugs are not (Yamada et al., 1994), anxiolytic (Bradley et al., 2007), anti- without side effects. Therefore, the search for regularly eaten depressive (Seol et al., 2010), and anticonflict effects (Umezu foods with an antidepressant activity seems to be an essential et al., 2006), have also been reported. approach to finding an effective antidepressant treatment On the other hand, lavender is also used as a tea infusion without side effects. (i.e., aqueous extracts) to treat restlessness, insomnia, and nervous disorders of the stomach and intestines (Blumenthal *To whom correspondence should be addressed. et al., 1998). Furthermore, lavender contains aqueous phe- E-mail: [email protected] nolic constituents, such as hydroxycinnamic acids and fla- 474 A. Kageyama et al. vone glycosides (Harborne and Williams, 2002), which have solt et al., 1977, Kageyama et al., 2010). Briefly, 35 male been associated with the antioxidant activities of Lamiaceae rats were divided into five groups (seven rats per group): one plants including lavender (Zheng and Wang, 2001; Dorman group served as the control, another group received imip- et al., 2004; Shan et al., 2005; Torras-Claveria et al., 2007). ramine (30 mg/kg body weight), and the other three groups However, in contrast to the essential oil, few studies have in- received LAE at three different doses (857, 1714, and 3428 vestigated the medicinal properties of the aqueous extracts of mg/kg). The rats were placed in an acrylic cylinder (45 cm lavender. × 19.2 cm i.d.) filled with water at 25 ± 1℃ to a depth of The purpose of the present study was to examine the 17 cm for 15 min without any treatment (pre-test session). antidepressant-like effects of an aqueous extract of lavender Twenty-four hours later, the rats were exposed to the same (LAE) by using the forced swimming test (FST), the most conditions for 5 min (test session). Between the pre-test and widely used animal model of depression. test sessions, the test solutions were orally administered three times, the first just after the pre-test session, then 5 h before Material and Methods the test session, and finally 1 h before the test session. Time Animals 6-week-old male Wistar rats (110 − 130 g) spent by the rats in active movement and immobility was re- were obtained from Japan SLC (Shizuoka, Japan). The rats corded during the 5-min test session by a video camera. The were group-housed (seven rats/cage; cage size: 420 × 263 × tapes were evaluated by observers not informed of the treat- 199 mm height) in a room at a constant temperature (23 ± ment that each animal had received. After each test, the tank 1℃) and humidity (50 ± 5%), and a 12-hour light-dark cycle was refilled with fresh water. (light: 7:00 − 19:00). Controlled purified diets were prepared Open field test (OFT) procedure The locomotor activ- according to CE-2 (CLEA Japan, Inc., Shizuoka, Japan). The ity in a novel environment was recorded employing the OFT. animals consumed the diets and tap water ad libitum. All Fourteen male rats were divided into two groups (seven rats animals were handled daily for 7 days to familiarize them per group): one group served as the control and the other with the experimental equipment and to minimize nonspe- group received LAE at a dose of 3428 mg/kg. The test solu- cific stress responses during the experiment. Behavioral tests tions were orally administered three times, i.e., 24, 5, and 1 were performed between 13:00 and 17:00 h. A different set h before the test. The arena was a circular black box (70 cm of animals was used for each behavioral test. All experimen- diameter and 40 cm height). The test was begun by placing tal procedures were in accordance with the Guidelines of the the animals at the center of the arena. The test used a CCD University of Shizuoka for the Care and Use of the Labora- camera (2 frames/s) and a computer video tracking system tory Animals, based on those of the American Association Image OF (O'hara & Co., Ltd., Tokyo, Japan), modified soft- for Laboratory Animal Science. ware based on the public domain NIH Image program (i). Chemicals Imipramine hydrochloride was purchased Locomotion (cm) and percent center (%) were analyzed for from Nacalai Tesque (Kyoto, Japan). (-)-Linalool and rosma- 5 min. Defecation was estimated by counting the number of rinic acid were purchased from Sigma-Aldrich Japan (Tokyo, fecal boluses deposited in the arena. Japan). Apigenin was purchased from Wako Pure Chemi- High performance liquid chromatography (HPLC) An cal Industries (Osaka, Japan). tert-Butylhydroquinone was Agilent 1200 Series HPLC system equipped with a diode purchased from Kanto Chemicals (Tokyo, Japan). All other array detector (Agilent Technologies, Palo Alto, CA) and reagents were of the highest grade commercially available. a Capcell Pak C18 MG column (250 mm × 4.6 mm i.d.; Preparation of LAE One hundred grams of dried lav- particle size, 5 µm; Shiseido, Tokyo, Japan) were used. The ender (Lavandula angustifolia Mill.) flowers were extracted operating conditions were as follows: column oven tem- with 2 L of distilled water at 90℃ for 1 h. The extract sepa- perature, 40℃; mobile phase, a linear gradient from 100% rated from the residue was evaporated in vacuo and lyophi- solvent A (water, pH 2.5 adjusted with phosphoric acid/ace- lized to give 25 g of LAE. The LAE was stored in a desicca- tonitrile 90:10) to 100% solvent B (acetonitrile) in 30 min, tor at 4℃ until needed. held at 100% B for 5 min; flow rate, 1 mL/min; and injection Treatments LAE and imipramine hydrochloride were volume, 1 µL. The samples were filtered though a 0.45-µm dissolved in distilled water just before use. All solutions PTFE membrane filter (DISMIC-13HP, Toyo Roshi, Tokyo, were orally administered at a volume of 1.0 mL/100 g body Japan) prior to injection. Wavelengths of 210, 310, and 350 weight of the rat. The control group only received distilled nm were used for the quantification of linalool, rosmarinic water. acid, and apigenin aglycone, respectively, and the UV spectra FST procedure The FST was performed according to (wavelengths from 200 to 400 nm) plus retention time were the method of Porsolt et al., with some modifications (Por- used for their identification. Antidepressant-like Effects of Lavender 475

Determination of total apigenin glycosides in LAE To result of imipramine was not significant, the analysis was determine the total concentration of apigenin glycosides in stopped so that the overall FWER did not exceed a level of LAE, hydrolysis was performed by a method of Hertog et 0.05 (two-tailed). In the case of data from the OFT, the dif- al. (1992) with some modifications. Two milliliters of 62.5% ference between the vehicle control and LAE at a dose of aqueous methanol containing 25 mg of LAE and 4 mg of 3428 mg/kg was examined by the Student’s t-test at a signifi- TBHQ were mixed with 0.5 mL of 10 M HCl. After refluxing cance level of 0.05 (two-tailed). Prior to the Student’s t-test at 90℃ for 4 h, the solution was cooled to room temperature and Williams’ test, homogeneity of variance was checked us- and extracted by adding ethyl acetate (5 mL) and water (3 ing F-test and Bartlett’s test, respectively, and a significance mL). The ethyl acetate layer was washed twice with water (5 level of 0.05 was used for each test. mL), dried over sodium sulfate, and subjected to HPLC analysis. The total concentration of apigenin glycosides in LAE Results was calculated by the difference between the concentration Immobility time in the FST LAE (857, 1714, and 3428 of apigenin aglycone before and after hydrolysis. mg/kg) and imipramine (30 mg/kg) were orally administered Statistical analysis Data are expressed as mean ± stan- to the rats 24, 5, and 1 h before the test. No deaths or evident dard error of the mean (SEM) and were analyzed using Mi- toxic effects, such as vomiting, diarrhea or abnormal behav- crosoft Excel 2007 (Microsoft Japan, Tokyo, Japan) with the ior, were observed in any of the groups. Figure 1 shows the add-in software Statcel 3 (OMS Publishing, Saitama, Japan). effects of imipramine and LAE on the immobility time in the For the analysis of the immobility time in the FST, a fixed- FST. The mean immobility time and SEM in rats treated with sequence testing procedure (Dmitrienko et al., 2005) was ad- imipramine was 154 ± 16 s, which was significantly lower opted to control the family-wise type I error rate (FWER) for (t = 3.13, df = 12, p = 0.009) than that of the vehicle control the two steps of statistical analysis, which were performed rats (236 ± 21 s), indicating that the test had sufficient sen- in the following pre-specified order: first, the difference be- sitivity to detect antidepressant-like action of the positive tween the vehicle control and imipramine (a positive control) control drug at a dose previously reported to be effective in was examined by the Student’s t-test at a significance level of the FST in rats (Butterweck et al., 2001). On the other hand, 0.05 (two-tailed) in order to assess the validity of the experi- the immobility time in rats treated with LAE at the low, ment: then, if the result of imipramine was significant, the ef- medium, and high doses was 242 ± 18, 234 ± 12, and 178 ± fect of three different doses of LAE compared to the vehicle 23 s, respectively, indicating a dose-dependent reduction of control was examined by Williams’ test (Williams, 1972) at immobility time. Williams’ test revealed a significant effect a significance level of 0.025 (one-tailed) on the assumption of LAE at a dose of 3428 mg/kg as compared to the vehicle that a dose-dependent effect exists. On the other hand, if the control group (Williams’ test statistic of 2.17, four groups, df

300

250 )

sec * (

200

e ＃ m i T

y 150 t ili ob 100 mm I

0 Control Imipramine 857 1714 3428 LAE (mg/kg) Fig. 1. Effects of LAE on the FST immobility time in rats. Data are expressed as mean ± SEM (n = 7). A fixed-sequence procedure was adopted for statistical analysis. #p < 0.05 (two-tailed) vs. control by Student’s t-test. *p < 0.025 (one-tailed) vs. control by Williams’ test. 476 A. Kageyama et al.

Table 1. Effects of LAE on the OFT in rats.

Control LAE (3428 mg/kg) t a (df b = 12) p (two-tailed) Locomotion cm 1909 ± 67 1647 ± 147 1.63 0.13 Center % 27 ± 2 24 ± 2 1.15 0.27 Defecation Number 0.8 ± 0.6 0.5 ± 0.5 0.24 0.82

Each value is the mean ± SEM (n = 7). a Student’s t-statistic. b Degrees of freedom.

mAu

100

80 A Linalool

0 2.5 5 7.5 10 12.5 15 17.5 20 22.5 min mAu

100

80 B

0 2.5 5 7.5 10 12.5 15 17.5 20 22.5 min

Fig. 2. HPLC chromatograms of LAE before (A) and after (B) the evaporation-lyophilization process. A wavelength of 210 nm was used for recording the chromatograms.

= 24, p < 0.025 [one-tailed]). The percent of reduction in the Table 2. Concentation of rosmarinic acid, apigenin, and immobility time in rats treated with LAE (3428 mg/kg) was apigenin glycosides in LAE. 25%, which was comparable to some extent to that of the Concentration Compound positive control imipramine (35%). (μmol/g dry weight) Locomotor activity in the OFT LAE (3428 mg/kg) was Rosmarinic acid 37.1 orally administered to the rats 24, 5, and 1 h before the test. Apigenin glycosides 5.5 Table 1 shows the effects of LAE on the locomotion (cm), Apigenin < 0.1 time spent in the center of the arena (%), and number of def- ecations in the OFT, among which the last two parameters are used as indicators of anxiolytic effects (Prut and Bel- cess was determined to be 24.2 mg/g on a dry weight basis, zung, 2003). The administration of LAE did not significantly whereas linalool was not detected after the evaporation- affect any of these parameters in the OFT but reduced the lyophilization process (Fig. 2). locomotion (p = 0.13) by 14% (95% confidence interval: −5 Concentration of rosmarinic acid, apigenin, and apigenin to +32%). This might suggest a mild sedative action of LAE. glycosides in LAE The concentration of rosmarinic acid, A number of antidepressants including imipramine have also apigenin, and apigenin glycosides in LAE was determined as been reported to decrease the open field activity in rats at shown in Table 2. The results showed that apigenin presents doses that exert antidepressant action in the FST (Porsolt et mostly as glycosides in LAE. al., 1978). Concentration of linalool in LAE before and after the Discussion evaporation-lyophilization process The concentration of The FST (Porsolt et al., 1977) is the most widely used linalool in LAE before the evaporation-lyophilization pro- animal model for screening potential antidepressant activity. Antidepressant-like Effects of Lavender 477

The immobility displayed by rodents when subjected to an FST (Seol et al., 2010). These controversial results might unavoidable stress such as forced swimming was assumed be due to differences in the mode of administration. On the to be a state of despair, in which the animal has learned that other hand, the oral administration of lavender tincture (an escape is impossible (Porsolt et al., 1977). Additionally, the aqueous ethanolic extract) was reported to enhance the thera- immobility time in the FST has been shown to be reduced peutic effect of imipramine in depressed patients (Akhondza- by treatment with known antidepressant drugs (Porsolt et al., deh et al., 2003), whereas the active constituents responsible 1977; Porsolt et al., 1978). Moreover, a strong correlation for this adjuvant effect have not been clarified. In the present has been reported between the clinical efficacy of the antide- study, linalool, the main essential oil constituent of lavender, pressant drugs and their potency in the FST (Willner, 1984). was completely removed by the evaporation-lyophilization Nevertheless, psychostimulants, such as d-amphetamine and process (Fig. 2). Therefore, the antidepressant-like effects of caffeine, have been reported to decrease the immobility time LAE can be attributed to its non-volatile constituents. Laven- in the FST apparently due to motor stimulation rather than der has been reported to contain non-volatile phenolic com- to persistent attempts to escape (Porsolt et al., 1977; Porsolt pounds such as hydroxycinnamic acids and flavone glyco- et al., 1978). These false-positive results can be excluded by sides (Harborne and Williams, 2002), which can be extracted an OFT, in which clinically effective antidepressant drugs re- by hot water and were possibly present in the LAE used in duced the locomotor activity, but psychostimulants markedly this study. Among the phenolic compounds previously found increased it (Porsolt et al., 1978). In the present study, the in lavender, apigenin, which is mostly present as a glycoside oral administration of LAE to rats at a dose of 3428 mg/kg in lavender (Harborne and Williams, 2002), and rosmarinic showed a significant reduction of the immobility time during acid have been reported to have an antidepressant-like activ- the FST, the effect of which was comparable to that of the ity in the FST (Takeda et al., 2002a; Takeda et al., 2002b; synthetic antidepressant imipramine at a dose of 30 mg/kg Nakazawa et al., 2003; Yi et al., 2008). (Fig. 1). In addition, the same dose of LAE did not affect the In order to estimate the contribution of rosmarinic acid locomotor activity during the OFT (Table 1), indicating that and apigenin glycosides to the antidepressant-like effects the antidepressant-like effect of LAE in the FST cannot be of LAE, the concentration of these compounds in LAE was attributed to motor stimulation. determined (Table 2). From these data, the amount of ros- In this study, an antidepressant-like effect of LAE in rats marinic acid in the effective dose of LAE was calculated to was demonstrated after acute treatment (three administra- be 127 μmol/kg (37.1 µmol/g × 3.428 g/kg body weight). tions in 24 h) at a relatively high dose of 3428 mg/kg. On This value is greater than 99.1 μmol/kg, a reported oral dose the other hand, clinical efficacy of antidepressants such as acutely effective in the FST in mice (Takeda et al., 2002b). imipramine in humans becomes evident after chronic treat- Therefore, rosmarinic acid seems to be one of the active ment for 2 − 3 weeks, even though most of them are acutely compounds for the antidepressant-like effects of LAE. In effective in the FST at doses 10- to 100-fold greater than the same way, the total amount of apigenin glycosides in the those used in clinical practice on a milligram per kilogram effective dose of LAE was calculated to be 18.9 μmol/kg. per day basis (Borsini and Meli, 1988; Detke et al., 1997). This value is less than 37.0 μmol/kg, a reported oral dose of In addition, it has been demonstrated that low doses of the apigenin aglycone effective in the FST in mice, not acutely antidepressants desipramine and fluoxetine that were inactive but after 14 days treatment (Yi et al., 2008). In addition, in the FST after acute treatment became active after chronic bioavailability of apigenin glycosides in rats might be lower treatment (Detke et al., 1997). These findings suggest that than that of apigenin aglycone, because intestinal permeabil- LAE at doses less than 3425 mg/kg might be effective in the ity of apigenin 7-O-glucoside, an apigenin glycoside found FST after chronic treatment and this time course might agree in lavender (Harborne and Williams, 2002), was reported more with possible use of LAE in humans. Thus, chronic to be much lower than that of apigenin aglycone in both the treatment remains to be studied in order to fully understand caco-2 cell culture model and perfused rat intestinal model the efficacy of LAE in the FST. (Liu and Hu, 2002). Therefore, roles of apigenin glycosides Specific constituents responsible for the antidepressant- in the efficacy of LAE after acute treatment seem to be less like effects of LAE are still unclear. It has been reported that important. Compounds other than rosmarinic acid and api- vapor inhalation of the lavender essential oil did not affect or genin glycosides remain to be investigated for their possible even increase the immobility time in the FST presumably due roles in the antidepressant-like effects of LAE. to its sedative effect (Komiya et al., 2006; Lim et al., 2005). In conclusion, LAE showed an antidepressant-like effect In contrast, intraperitoneal administration of the lavender es- in the FST after acute treatment and rosmarinic acid was sential oil was reported to reduce the immobility time in the suggested to be one of the active ingredients in LAE. Fur- 478 A. Kageyama et al. ther studies are needed to clarify the efficacy of LAE after Dmitrienko, A., Molenberghs, G., Chuang-Stein, C. and Offen, chronic treatment and contribution of other compounds to W. (2005). “Analysis of Clinical Trials Using SAS: A Practical the antidepressant-like effects of LAE. Guide.” SAS Institute, Cary, NC. Dorman, H.J., Bachmayer, O., Kosar, M. and Hiltunen, R. (2004). Acknowledgements This work was supported by the Laboratory of Antioxidant properties of aqueous extracts from selected lamia- Nutritional Biochemistry members at the University of Shizuoka, ceae species grown in Turkey. J. Agric. Food Chem., 52, 762- and also supported in part by the Global COE program in Japan. 770. Harborne, J.B. and Williams, C.A. (2002). Phytochemistry of the References genus Lavandula. In “Lavender: the genus Lavandula,” ed. by M. Akhondzadeh, S., Kashani, L., Fotouhi, A., Jarvandi, S., Mobaseri, Lis-Balchin. Taylor & Francis, London, pp. 86-99. M., Moin, M., Khani, M., Jamshidi, A.H., Baghalian, K. and Hertog, M.G.L., Hollman, P.C.H. and Venema, D.P. (1992). Opti- Taghizadeh, M. (2003). Comparison of Lavandula angustifolia mization of a quantitative HPLC determination of potentially an- Mill. tincture and imipramine in the treatment of mild to moder- ticarcinogenic flavonoids in vegetables and fruits. J. Agric. Food ate depression: a double-blind, randomized trial. Prog. Neuropsy- Chem., 40, 1591-1598. chopharmacol. Biol. Psychiatry, 27, 123-127. Kageyama, A., Sakakibara, H., Zhou, W., Yoshioka, M., Ohsumi, M., Andrade, L., Caraveo-Anduaga, J.J., Berglund, P., Bijl, R.V., De Shimoi, K. and Yokogoshi, H. (2010). Genistein regulated seroto- Graaf, R., Vollebergh, W., Dragomirecka, E., Kohn, R., Keller, M., nergic activity in the hippocampus of ovariectomized rats under Kessler, R.C., Kawakami, N., Kiliç, C., Offord, D., Ustun, T.B. forced swimming stress. Biosci. Biotechnol. Biochem., 74, 2005- and Wittchen, H.U. (2003). The epidemiology of major depres- 2010. sive episodes: results from the International Consortium of Psy- Komiya, M., Takeuchi, T. and Harada, E. (2006). Lemon oil vapor chiatric Epidemiology (ICPE) Surveys. Int. J. Method Psychiatr. causes an anti-stress effect via modulating the 5-HT and DA ac- Res., 12, 3-21. tivities in mice. Behav. Brain Res., 172, 240-249. Blumenthal, M., Busse, W.R., Goldberg, A., Gruenwald, J., Hall, Kuroda, K., Inoue, N., Ito, Y., Kubota, K., Sugimoto, A., Kakuda, T., Riggins, C.W. and Rister, R.S. (eds.) (1998). “The complete T. and Fushiki, T. (2005). Sedative effects of the jasmine tea odor German Commission E monographs: Therapeutic guide to herbal and (R)-(-)-linalool, one of its major odor components, on auto- medicine.” American Botanical Council, Austin. nomic nerve activity and mood states. Eur. J. Appl. Physiol., 95, Borsini, F. and Meli, A. (1988). Is the forced swimming test a suit- 107-114. able model for revealing antidepressant activity? Psychopharma- Lim, W.C., Seo, J.M., Lee, C.I., Pyo, H.B. and Lee, B.C. (2005). cology, 94, 147-160. Stimulative and sedative effects of essential oils upon inhalation Bradley, B.F., Starkey, N.J., Brown, S.L. and Lea, R.W. (2007). in mice. Arch. Pharm. Res., 28, 770-774. Anxiolytic effects of Lavandula angustifolia odour on the Mon- Lis-Balchin, M. (2006). “Aromatherapy science: a guide for health- golian gerbil elevated plus maze. J. Ethnopharmacol., 111, 517- care professionals.” Pharmaceutical Press, London. 525. Liu, Y. and Hu, M. (2002). Absorption and metabolism of flavo- Buchbauer, G., Jirovetz, L., Jäger, W., Dietrich, H. and Plank, C. noids in the caco-2 cell culture model and a perused rat intestinal (1991). Aromatherapy: evidence for sedative effects of the essen- model. Drug Metab. Dispos., 30, 370-377. tial oil of lavender after inhalation. Z. Naturforsch. C, 46, 1067- Nakazawa, T., Yasuda, T., Ueda, J. and Ohsawa, K. (2003). Antide- 1072. pressant-like effects of apigenin and 2,4,5-trimethoxycinnamic Buchbauer, G., Jirovetz, L., Jäger, W., Plank, C. and Dietrich, H. acid from Perilla frutescens in the forced swimming test. Biol. (1993). Fragrance compounds and essential oils with sedative ef- Pharm. Bull., 26, 474-480. fects upon inhalation. J. Pharm. Sci., 82, 660-664. Porsolt, R.D., Anton, G., Blavet, N. and Jalfre, M. (1978). Behav- Butterweck, V., Nishibe, S., Sasaki, T. and Uchida, M. (2001). Anti- ioural despair in rats: a new model sensitive to antidepressant depressant effects of Apocynum venetum leaves in a forced swim- treatments. Eur. J. Pharmacol., 47, 379-391. ming test. Biol. Pharm. Bull., 24, 848-851. Porsolt, R.D., Le Pichon, M. and Jalfre, M. (1977). Depression: a Detke, M.J., Johnson, J. and Lucki, I. (1997). Acute and chronic an- new animal model sensitive to antidepressant treatments. Nature, tidepressant drug treatment in the rat forced swimming test model 266, 730-732. of depression. Exp. Clin. Psychopharmacol., 5, 107-112. Prut, L. and Belzung, C. (2003). The open field as a paradigm to Diego, M.A., Jones, N.A., Field, T., Hernandez-Reif, M., Schan- measure the effects of drugs on anxiety-like behaviors: a review. berg, S., Kuhn, C., McAdam, V., Galamaga, R. and Galamaga, M. Eur. J. Pharmacol., 463, 3-33. (1998). Aromatherapy positively affects mood, EEG patterns of Seol, G.H., Shim, H.S., Kim, P.J., Moon, H.K., Lee, K.H., Shim, alertness and math computations. Int. J. Neurosci., 96, 217-224. I, Suh, S.H. and Min, S.S. (2010). Antidepressant-like effect of Antidepressant-like Effects of Lavender 479

Salvia sclarea is explained by modulation of dopamine activities tification of its active constituents. Pharmacol. Biochem. Behav., in rats. J. Ethnopharmacol., 130, 187-190. 85, 713-721. Shan, B., Cai, Y.Z., Sun, M. and Corke, H. (2005). Antioxidant ca- Williams, D.A. (1972). The comparison of several dose levels with pacity of 26 spice extracts and characterization of their phenolic a zero dose control. Biometrics, 28, 519-531. constituents. J. Agric. Food Chem., 53, 7749-7759. Willner, P. (1984). The validity of animal models of depression. Takeda, H., Tsuji, M., Inazu, M., Egashira, T. and Matsumiya, T. Psychopharmacology (Berl.), 83, 1-16. (2002a). Rosmarinic acid and caffeic acid produce antidepres- Yamada, K., Mimaki, Y. and Sashida, Y. (1994). Anticonvulsive ef- sive-like effect in the forced swimming test in mice. Eur. J. Phar- fects of inhaling lavender oil vapour. Biol. Pharm. Bull., 17, 359- macol., 449, 261-267. 360. Takeda, H., Tsuji, M., Matsumiya, T. and Kubo, M. (2002b). Identi- Yi, L.T., Li, J.M., Li, Y.C., Pan, Y., Xu, Q. and Kong, L.D. (2008). fication of rosmarinic acid as a novel antidepressive substance in Antidepressant-like behavioral and neurochemical effects of the the leaves of Perilla frutescens Britton var. acuta Kudo (Perillae citrus-associated chemical apigenin. Life Sci., 82, 741-751. Herba). Nihon Shinkei Seishin Yakurigaku Zasshi, 22, 15-22. Zheng, W. and Wang, S.Y. (2001). Antioxidant activity and pheno- Torras-Claveria, L., Jauregui, O., Bastida, J., Codina, C. and Vilado- lic compounds in selected herbs. J. Agric. Food Chem., 49, 5165- mat, F. (2007). Antioxidant activity and phenolic composition of 5170. Lavandin (Lavandula x intermedia Emeric ex Loiseleur) waste. J. Agric. Food Chem., 55, 8436-8443. URL Cited Umezu, T., Nagano, K., Ito, H., Kosakai, K., Sakaniwa, M. and i) http://rsb.info.nih.gov/nih-image/ (July 15,2011) Morita, M. (2006). Anticonflict effects of lavender oil and iden-